PONE-D-19-35054

Abrogation of pathogenic attributes in drug resistant Candida auris strains by farnesol

PLOS ONE

Dear Dr. Ahmad

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Due to the importance of the multi-drug resistant strain Candida auris, this manuscript is of interest. However, there are some issues that need to be addressed. First the material and methods re strains and PCR need to be expanded upon. Since C. auris is reported to demonstrate more MDR than other strains, there should be a short discussion on the relative sensitivity between C. auris and other Candida strains to farnesol. Finally both reviewers have noted other minor points that should be addressed. Please address both reviewers.

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PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Manuscript describes the impact of farnesol on Candida auris strains, believed to be all belonging to the South African-clade as they are all from South Africa (but there is no proper ID provided, so some isolates can be from Asia or South America). A subset is chosen, but it is unclear on which grounds, to run the experiments on. The manuscript itself seems be hastily written and this needs more attention. Another flaw is that statements are made that (for example) a RT-qPCR method for the expression of a large number of genes is used as published before... but only part of the genes were used in the previously published study. On the next page authors provide primers for the missing genes, but this is not a proper Material & Methods description. For most of the experiments a cell suspension is used, it is unknown how the authors counted the cells and if this is based on a McFarland range or that a cell counter was used. See more comments in the attached file (up to page 14 as the reminder of the document does not contain that many comments; except that the references needs to be checked to be in the correct style).

Reviewer #2: This study investigates the effect of a quorum-sensing molecule (farnesol) on the emerging yeast Candida auris, which is particularly virulent and often resistant to the usual antifungal agents. For these reasons, the data reported in this study is worth considering for publication in PLOS ONE. The pre-print of this manuscript has been deposited on the BioRxiv server.

The submitted manuscript is well written. The authors have carefully chosen different techniques to analyze the effect of the quorum-sensing molecule (farnesol) on C. auris. The authors have combined the description and interpretation of the results for each of the techniques considered. This makes the manuscript clear and easy to read. However, a general discussion/conclusion would be essential to discuss the sensitivity of C. auris to farnesol observed in this study, with the data available in the literature for other Candida’s. Thus, the authors could emphasize that their strains of C. auris sometimes present a higher MIC for farnesol than that observed by other authors in other species of Candida.

Below you will find some remarks that should be clarified for a better reading/understanding.

1 °) Introduction and/or general conclusion. Authors should justify the interest of farnesol to inhibit the growth of C. auris compared to other quorum-sensing molecules (such as tyrosol) or compared to molecules belonging to other physiological mechanisms of defense.

2 °) Material and methods. C. albicans SC5314 is well known as a reference strain from a library. The authors should further specify the status of their C. auris strains: reference strains from a bacterial library or wild strains well characterized by a National Institute? If the authors consider the strains studied as isolates from their countries, is it worth mentioning the internal laboratory identification number? This could contribute to lightening the text.

3 °) Material and methods. Lines 101-103: The origin of farnesol should be mentioned as well as the concentration of the stock solution. Are farnesol concentrations (from 250 to 0.48 mM) indicative of reactant concentration or final concentration in the assay tube/well? Line 132: "500 to 0.488 mM “ rather than "500 mM to 0.488 mM”.

Minor remarks.

Line 55-56: Abbreviations of amphotericin (AmB) and fluconazole (FLZ) should be clearly indicated in parentheses after the first mention of each in the text. “…resistant to azoles as fluconazole (FLZ) and …unaffected to polyenes as amphotericin (AmB). …”

Line 114: Revise the sentence: “… the lowest concentration of (?) AmB farnesol that …

Line 192: Choose 20h or 1200 min: only one of both is enough.

Line 290: Clarify “pre-incubation (0h)” statement. Does it well mean the absence of pre-incubation?

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: PONE-D-19-35054_REVIEW.pdf

Abrogation of pathogenic attributes in drug resistant Candida auris strains by farnesol

PONE-D-19-35054R1

Dear Dr. Ahmad,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

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With kind regards,

Joy Sturtevant

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: (No Response)

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: (No Response)

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: (No Response)

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: (No Response)

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: no additional questions/comments as previously raised ones were addressed, the manuscript has been thoroughly revised.

Reviewer #2: After re-consideration by the authors, the data reported in this manuscript deserves to be published in PLOS-ONE.

The authors conscientiously took into account the concerns and remarks of the reviewers, in particular concerning the description of the procedures and the origin with the characterization of the strains tested. The writing has been greatly improved. It remains 3 minor typing mistakes:

"C. auris" (l.139), "C. albicans" (l.141-142) and "Candida" (l. 145) must be in italic.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No